摘要
目的探讨硫化氢供体硫氢化钠对过氧化氢诱导心肌细胞氧化应激损伤的保护机制。方法采用胶原酶消化分离培养SD大鼠乳鼠心肌细胞,过氧化氢诱导建立氧化应激模型。经硫氢化钠和LY294002预处理探讨硫化氢对过氧化氢诱导心肌细胞损伤的保护作用,观察心肌细胞形态学变化,检测乳酸脱氢酶、丙二醛的含量和超氧化物歧化酶的活性,Western blot检测凋亡相关蛋白表达,流式细胞术检测心肌细胞的凋亡。结果过氧化氢可引起乳鼠心肌细胞乳酸脱氢酶、丙二醛含量升高,Akt磷酸化水平下降,Bcl-2/Bad比值降低,细胞凋亡率升高(P<0.05);硫氢化钠预处理能减轻过氧化氢引起的心肌细胞损伤,乳酸脱氢酶、丙二醛含量下降,Akt磷酸化水平升高,Bcl-2/Bad比值升高,细胞凋亡率下降(P<0.05);PI3K/Akt特异性阻断剂LY294002能部分减轻硫氢化钠的保护作用(P<0.05)。结论硫氢化钠预处理能使Akt的磷酸化水平升高,Bcl-2/Bad比值升高,在一定程度上减轻过氧化氢诱导的心肌细胞损伤及凋亡,硫化氢可能通过激活PI3K/Akt信号转导通路发挥保护作用。
Aim To study the cardioprotective mechanism of hydrogen sulfide（H2S） on oxidative stress induced by hydrogen peroxide. Methods Primary-cultured cardiomyocytes were obtained from neonatal rats,an oxidative stress injury model was established by exposing the cells to H2O2 for 2 h.To study the cardioprotective mechanism of hydrogen sulfide（H2S） on oxidative stress induced by hydrogen peroxide,the cells were pre-incubated with ly294002 for 30 min.The morphology change of cardiomyocytes was observed by inverted phase contrast microscope,the activities of lactate dehydrogenase（LDH） and superoxide dismutase（SOD）,and content malondialdehyde（MDA） were determined,the activation of the PI3K/Akt signal pathway was observed by Western blotting,the cardiomyocytes apoptosis was detected by AnnexinV/PI flow cytometry. Results After exposing to H2O2 for 2 h,the release of LDH,MDA and apoptosis were increased,the activities of SOD and ratio of Bcl-2/Bad were decreased（P〈0.05）.After pretreated with sodium hydrosulfide（NaHS） for 30 mins,the injury effect was attenuated,moreover,the phosphorylation of Akt,Bad and ratio of Bcl-2/Bad were upregulated.LY294002 could block the protection effect of NaHS partly. Conclusion H2S can attenuate hydrogen peroxide-induced rat neonatal cardiomyocytes injury,which may involve the PI3K/Akt pathway.
出处
《中国动脉硬化杂志》
CAS
CSCD
北大核心
2011年第4期310-314,共5页
Chinese Journal of Arteriosclerosis
